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Basal medium composition and serum or serum replacement concentration influences on the maintenance of murine embryonic stem cells

机译:基础培养基组成和血清或血清替代浓度对鼠胚胎干细胞维持的影响

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摘要

The expansion of stem cell numbers while retaining their developmental properties is a bioprocess challenge. We compared the growth rates and embryoid body (EB) formation yields of R1 and EFC murine embryonic stem cells (mESC) cultured in two basal media (DMEM or DMEM:F12) with additions of 1.7–15% fetal bovine serum (FBS) or serum replacer (KOSR). Whereas the basal medium or KOSR dose did not have a significant effect on growth rate for either cell line, increasing doses of KOSR had a significant negative effect on the EB yield of EFC cells. Use of DMEM:F12 and increasing doses of FBS independently and significantly increased the growth rate for both cell lines. DMEM:F12 also significantly increased EB yields for both cell lines. The results show that use of DMEM:F12 and several-fold lower than conventional concentrations of KOSR can efficiently support maintenance of mESC and that KOSR should be dose as well as lot optimized.
机译:干细胞数量的扩展同时保持其发育特性是生物过程的挑战。我们比较了在两种基础培养基(DMEM或DMEM:F12)中培养的R1和EFC鼠类胚胎干细胞(mESC)的生长速率和胚状体(EB)生成量,并添加了1.7–15%胎牛血清(FBS)或血清替代品(KOSR)。尽管基础培养基或KOSR剂量对任一细胞系的生长速率均无显着影响,但增加剂量的KOSR对EFC细胞的EB产量具有显着的负面影响。 DMEM:F12的使用和FBS剂量的独立增加,均显着提高了两种细胞系的生长速率。 DMEM:F12还显着提高了两种细胞系的EB产量。结果表明,使用DMEM:F12且比常规KOSR浓度低几倍可以有效地支持mESC的维持,并且KOSR应该在剂量和批次上进行优化。

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